Is there have Finished kit for Coupling or Electron Flow measurement in isolated mitochondrial with XFe24 or XFe96?

Hi LD,

Thank you for contacting Cell Analysis Technical Support. Unfortunately, we do not have a kit for Coupling or Electron Flow, but the below paper does a great job with reference materials, including part numbers and manufacturer. 

Measuring Mitochondrial Function in Permeabilized Cells Using the Seahorse XF Analyzer or a Clark-Type Oxygen Electrode

Regarding when to add the MAS buffer, please see below from our Iso Mito App Note:

"XF Assay Preparation To minimize variability between wells, mitochondria were first diluted 10x in cold 1x MAS + substrate, then subsequently diluted to the needed concentration required for plating. Note that substrates included in the initial dilution, and is present during the centrifugation step."

Thank you and please let me know if you have any other questions!

Courtney

Courtney Nadeau Watts

Technical Support Scientist/Remote Engineer

Cell Analysis Products

Phone Contact: 800-227-9770 (Option 3 , option 8)

Email Contact: cellanalysis.support@agilent.com

Hi LD,

Thank you for contacting Cell Analysis Technical Support. Unfortunately, we do not have a kit for Coupling or Electron Flow, but the below paper does a great job with reference materials, including part numbers and manufacturer. 

Measuring Mitochondrial Function in Permeabilized Cells Using the Seahorse XF Analyzer or a Clark-Type Oxygen Electrode

Regarding when to add the MAS buffer, please see below from our Iso Mito App Note:

"XF Assay Preparation To minimize variability between wells, mitochondria were first diluted 10x in cold 1x MAS + substrate, then subsequently diluted to the needed concentration required for plating. Note that substrates included in the initial dilution, and is present during the centrifugation step."

Thank you and please let me know if you have any other questions!

Courtney

Courtney Nadeau Watts

Technical Support Scientist/Remote Engineer

Cell Analysis Products

Phone Contact: 800-227-9770 (Option 3 , option 8)

Email Contact: cellanalysis.support@agilent.com

Hi Courtney,

Thank you for your reply. Your reply has cleared up many of my doubts.

Different tissues may have different preferences for metabolic substrates, so are there any guidelines for the metabolic substrates of mitochondria sourced from different tissues for use in Seahorse assays? For instance, mitochondrial substrates from cardiac tissue or skeletal muscle tissue.

I look forward to hearing from you.

Best Regards,

LD

Hi LD,

You are very welcome! Table 25.2.1 in the paper above gives recommendations. You can also take a look at our Cell Publication Database to see what others have done as far as this.

Thank you and let me know if you have any other questions.

Courtney